g(m2)-ganglioside and Pheochromocytoma

Rat clonal pheochromocytoma PC12h cells were found to bind beta-galactosidase modified with specific glycosides. The enzyme modified with p-aminophenyl beta-D-glucoside was most effectively bound to the cells, followed by alpha-D-mannoside and alpha-D-glucoside. The binding was dependent on the number of PC12h cells, the incubation interval, and the pH; the maximal binding at 4 degrees C was obtained by incubation with 75 micrograms of cell protein for 15 min at pH 4.0. The binding proved to be a saturable and receptor-mediated process, and the apparent Km value and the maximal binding capacity of the cells with beta-D-glucosylated beta-galactosidase were 1.03 +/- 0.06 microM and 333 +/- 24 pmol/min/mg of protein, respectively. When the cells were cultured in the presence of nerve growth factor (NGF), GM1, GM2, and a ganglioside mixture, marked morphological differentiation was observed in the presence of NGF, and the specificity of the binding was also affected. By supplementation of NGF in the culture medium, the cells lost the selectivity of the glycoside binding, whereas cells cultured with GM1 supplement showed increased binding of the specific glycosides.

Topics: Adrenal Gland Neoplasms; Animals; beta-Galactosidase; Clone Cells; G(M1) Ganglioside; G(M2) Ganglioside; Galactosidases; Gangliosides; Glucosides; Glycosides; Glycosylation; Kinetics; Nerve Growth Factors; Pheochromocytoma; Rats; Tumor Cells, Cultured

The effects of ganglioside supplementation of culture medium on monoamine oxidase (MAO) type A and B activities in a rat clonal pheochromocytoma cell line, PC12h, were examined. The MAO activity in PC12h cells proved to be mainly due to type A MAO, and type B MAO activity was negligible. After supplementation of the culture medium with ganglioside GM1, the PC12 cells were found to express type B MAO activity after 4 days of culture, and the amount of type B activity increased with the number of days of culture. After 3 weeks of culture in the presence of GM1, type B activity was about 10% of the total, whereas in control cells type B MAO activity was only about 0.6% of the total. By kinetic analyses of type A and B MAO in PC12h cells after 3 weeks of culture, the increase of type B MAO activity was found to be due to the increase in amount of type B MAO; the Km values were almost the same and only the Vmax values were increased in the cells supplemented with GM1. Among gangliosides tested GM1 was the most effective in causing expression of type B MAO activity, whereas nerve growth factor was not effective. These results suggest that GM1 and other gangliosides may be involved in the expression of type B MAO in nerve cells and in the regulation of levels of the biogenic amines in the brain.

Topics: Adrenal Gland Neoplasms; Animals; Cell Line; Clorgyline; G(M1) Ganglioside; G(M2) Ganglioside; Kinetics; Kynuramine; Monoamine Oxidase; Nerve Growth Factors; Pheochromocytoma; Rats; Selegiline